Exposure protocol for ecotoxicity testing of microplastics and nanoplastics

Fazel Abdolahpur Monikh, Anders Baun, Nanna B. Hartmann, Raine Kortet, Jarkko Akkanen, Jae Seong Lee, Huahong Shi, Elma Lahive, Emilia Uurasjärvi, Nathalie Tufenkji, Korinna Altmann, Yosri Wiesner, Hans Peter Grossart, Willie Peijnenburg, Jussi V.K. Kukkonen

Research output: Contribution to journalArticlepeer-review

12 Scopus citations


Despite the increasing concern about the harmful effects of micro- and nanoplastics (MNPs), there are no harmonized guidelines or protocols yet available for MNP ecotoxicity testing. Current ecotoxicity studies often use commercial spherical particles as models for MNPs, but in nature, MNPs occur in variable shapes, sizes and chemical compositions. Moreover, protocols developed for chemicals that dissolve or form stable dispersions are currently used for assessing the ecotoxicity of MNPs. Plastic particles, however, do not dissolve and also show dynamic behavior in the exposure medium, depending on, for example, MNP physicochemical properties and the medium’s conditions such as pH and ionic strength. Here we describe an exposure protocol that considers the particle-specific properties of MNPs and their dynamic behavior in exposure systems. Procedure 1 describes the top-down production of more realistic MNPs as representative of MNPs in nature and particle characterization (e.g., using thermal extraction desorption-gas chromatography/mass spectrometry). Then, we describe exposure system development for short- and long-term toxicity tests for soil (Procedure 2) and aquatic (Procedure 3) organisms. Procedures 2 and 3 explain how to modify existing ecotoxicity guidelines for chemicals to target testing MNPs in selected exposure systems. We show some examples that were used to develop the protocol to test, for example, MNP toxicity in marine rotifers, freshwater mussels, daphnids and earthworms. The present protocol takes between 24 h and 2 months, depending on the test of interest and can be applied by students, academics, environmental risk assessors and industries.

Original languageEnglish
Pages (from-to)3534-3564
Number of pages31
JournalNature Protocols
Issue number11
StatePublished - Nov 2023
Externally publishedYes

Bibliographical note

Publisher Copyright:
© 2023, Springer Nature Limited.


This work received financial support from the UEF Water research program, which is jointly funded by the Saastamoinen Foundation, the Wihuri Foundation and the Olvi Foundation. The study was also partially funded by the European Union’s Horizon 2020 research and innovation program, via the projects PLASTICSFATE (Grant Agreement number 965367) and POLYRISK (Grant Agreement number 964766). N.T. acknowledges the Canada Research Chair program and the Killam Research Fellowship.

FundersFunder number
Wihuri Foundation
Itä-Suomen Yliopisto
Horizon 2020 Framework Programme964766, 965367
Canada Research Chairs
Saastamoisen säätiö


    Dive into the research topics of 'Exposure protocol for ecotoxicity testing of microplastics and nanoplastics'. Together they form a unique fingerprint.

    Cite this